Demethylase JMJD6 as a New Regulator of Interferon Signaling: Effects of HCV and Ethanol Metabolism

Murali Ganesan, Irina Tikhanovich, Shiva Shankar Vangimalla, Raghubendra Singh Dagur, Weimin Wang, Larisa Y Poluektova, Yimin Sun, David F Mercer, Dean Tuma, Steven A. Weinman, Kusum Kharbanda, Natalia A Osna

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Background & Aims: Alcohol-induced progression of hepatitis C virus (HCV) infection is related to dysfunction of innate immunity in hepatocytes. Endogenously produced interferon (IFN)α induces activation of interferon-stimulated genes (ISGs) via triggering of the Janus kinase–signal transducer and activator of transcription 1 (STAT1) pathway. This activation requires protein methyltransferase 1–regulated arginine methylation of STAT1. Here, we aimed to study whether STAT1 methylation also depended on the levels of demethylase jumonji domain-containing 6 protein (JMJD6) and whether ethanol and HCV affect JMJD6 expression in hepatocytes. Methods: Huh7.5-CYP (RLW) cells and hepatocytes were exposed to acetaldehyde-generating system (AGS) and 50 mmol/L ethanol, respectively. JMJD6 messenger RNA and protein expression were measured by real-time polymerase chain reaction and Western blot. IFNα-activated cells either overexpressing JMJD6 or with knocked-down JMJD6 expression were tested for STAT1 methylation, ISG activation, and HCV RNA. In vivo studies have been performed on C57Bl/6 mice (expressing HCV structural proteins or not) or chimeric mice with humanized livers fed control or ethanol diets. Results: AGS exposure to cells up-regulated JMJD6 expression in RLW cells. These results were corroborated by ethanol treatment of primary hepatocytes. The promethylating agent betaine reversed the effects of AGS/ethanol. Similar results were obtained in vivo, when mice were fed control/ethanol with and without betaine supplementation. Overexpression of JMJD6 suppressed STAT1 methylation, IFNα-induced ISG activation, and increased HCV-RNA levels. In contrast, JMJD6 silencing enhanced STAT1 methylation, ISG stimulation by IFNα and attenuated HCV-RNA expression in Huh7.5 cells. Conclusions: We conclude that arginine methylation of STAT1 is suppressed by JMJD6. Both HCV and acetaldehyde increase JMJD6 levels, thereby impairing STAT1 methylation and innate immunity protection in hepatocytes exposed to the virus and/or alcohol.

Original languageEnglish (US)
Pages (from-to)101-112
Number of pages12
JournalCMGH
Volume5
Issue number2
DOIs
StatePublished - 2018

Fingerprint

Hepacivirus
Interferons
Ethanol
Methylation
Acetaldehyde
Hepatocytes
Proteins
Betaine
RNA
Innate Immunity
Transcriptional Activation
Protein-Arginine N-Methyltransferases
Alcohols
STAT1 Transcription Factor
Viral Structural Proteins
Virus Diseases
Transducers
Genes
Arginine
Real-Time Polymerase Chain Reaction

Keywords

  • Alcohol
  • HCV
  • JMJD6
  • STAT1

ASJC Scopus subject areas

  • Hepatology
  • Gastroenterology

Cite this

Demethylase JMJD6 as a New Regulator of Interferon Signaling : Effects of HCV and Ethanol Metabolism. / Ganesan, Murali; Tikhanovich, Irina; Vangimalla, Shiva Shankar; Dagur, Raghubendra Singh; Wang, Weimin; Poluektova, Larisa Y; Sun, Yimin; Mercer, David F; Tuma, Dean; Weinman, Steven A.; Kharbanda, Kusum; Osna, Natalia A.

In: CMGH, Vol. 5, No. 2, 2018, p. 101-112.

Research output: Contribution to journalArticle

Ganesan, M, Tikhanovich, I, Vangimalla, SS, Dagur, RS, Wang, W, Poluektova, LY, Sun, Y, Mercer, DF, Tuma, D, Weinman, SA, Kharbanda, K & Osna, NA 2018, 'Demethylase JMJD6 as a New Regulator of Interferon Signaling: Effects of HCV and Ethanol Metabolism', CMGH, vol. 5, no. 2, pp. 101-112. https://doi.org/10.1016/j.jcmgh.2017.10.004
Ganesan, Murali ; Tikhanovich, Irina ; Vangimalla, Shiva Shankar ; Dagur, Raghubendra Singh ; Wang, Weimin ; Poluektova, Larisa Y ; Sun, Yimin ; Mercer, David F ; Tuma, Dean ; Weinman, Steven A. ; Kharbanda, Kusum ; Osna, Natalia A. / Demethylase JMJD6 as a New Regulator of Interferon Signaling : Effects of HCV and Ethanol Metabolism. In: CMGH. 2018 ; Vol. 5, No. 2. pp. 101-112.
@article{08824a1e682549b3b6ce188410b758a6,
title = "Demethylase JMJD6 as a New Regulator of Interferon Signaling: Effects of HCV and Ethanol Metabolism",
abstract = "Background & Aims: Alcohol-induced progression of hepatitis C virus (HCV) infection is related to dysfunction of innate immunity in hepatocytes. Endogenously produced interferon (IFN)α induces activation of interferon-stimulated genes (ISGs) via triggering of the Janus kinase–signal transducer and activator of transcription 1 (STAT1) pathway. This activation requires protein methyltransferase 1–regulated arginine methylation of STAT1. Here, we aimed to study whether STAT1 methylation also depended on the levels of demethylase jumonji domain-containing 6 protein (JMJD6) and whether ethanol and HCV affect JMJD6 expression in hepatocytes. Methods: Huh7.5-CYP (RLW) cells and hepatocytes were exposed to acetaldehyde-generating system (AGS) and 50 mmol/L ethanol, respectively. JMJD6 messenger RNA and protein expression were measured by real-time polymerase chain reaction and Western blot. IFNα-activated cells either overexpressing JMJD6 or with knocked-down JMJD6 expression were tested for STAT1 methylation, ISG activation, and HCV RNA. In vivo studies have been performed on C57Bl/6 mice (expressing HCV structural proteins or not) or chimeric mice with humanized livers fed control or ethanol diets. Results: AGS exposure to cells up-regulated JMJD6 expression in RLW cells. These results were corroborated by ethanol treatment of primary hepatocytes. The promethylating agent betaine reversed the effects of AGS/ethanol. Similar results were obtained in vivo, when mice were fed control/ethanol with and without betaine supplementation. Overexpression of JMJD6 suppressed STAT1 methylation, IFNα-induced ISG activation, and increased HCV-RNA levels. In contrast, JMJD6 silencing enhanced STAT1 methylation, ISG stimulation by IFNα and attenuated HCV-RNA expression in Huh7.5 cells. Conclusions: We conclude that arginine methylation of STAT1 is suppressed by JMJD6. Both HCV and acetaldehyde increase JMJD6 levels, thereby impairing STAT1 methylation and innate immunity protection in hepatocytes exposed to the virus and/or alcohol.",
keywords = "Alcohol, HCV, JMJD6, STAT1",
author = "Murali Ganesan and Irina Tikhanovich and Vangimalla, {Shiva Shankar} and Dagur, {Raghubendra Singh} and Weimin Wang and Poluektova, {Larisa Y} and Yimin Sun and Mercer, {David F} and Dean Tuma and Weinman, {Steven A.} and Kusum Kharbanda and Osna, {Natalia A}",
year = "2018",
doi = "10.1016/j.jcmgh.2017.10.004",
language = "English (US)",
volume = "5",
pages = "101--112",
journal = "Cellular and Molecular Gastroenterology and Hepatology",
issn = "2352-345X",
publisher = "Elsevier Inc.",
number = "2",

}

TY - JOUR

T1 - Demethylase JMJD6 as a New Regulator of Interferon Signaling

T2 - Effects of HCV and Ethanol Metabolism

AU - Ganesan, Murali

AU - Tikhanovich, Irina

AU - Vangimalla, Shiva Shankar

AU - Dagur, Raghubendra Singh

AU - Wang, Weimin

AU - Poluektova, Larisa Y

AU - Sun, Yimin

AU - Mercer, David F

AU - Tuma, Dean

AU - Weinman, Steven A.

AU - Kharbanda, Kusum

AU - Osna, Natalia A

PY - 2018

Y1 - 2018

N2 - Background & Aims: Alcohol-induced progression of hepatitis C virus (HCV) infection is related to dysfunction of innate immunity in hepatocytes. Endogenously produced interferon (IFN)α induces activation of interferon-stimulated genes (ISGs) via triggering of the Janus kinase–signal transducer and activator of transcription 1 (STAT1) pathway. This activation requires protein methyltransferase 1–regulated arginine methylation of STAT1. Here, we aimed to study whether STAT1 methylation also depended on the levels of demethylase jumonji domain-containing 6 protein (JMJD6) and whether ethanol and HCV affect JMJD6 expression in hepatocytes. Methods: Huh7.5-CYP (RLW) cells and hepatocytes were exposed to acetaldehyde-generating system (AGS) and 50 mmol/L ethanol, respectively. JMJD6 messenger RNA and protein expression were measured by real-time polymerase chain reaction and Western blot. IFNα-activated cells either overexpressing JMJD6 or with knocked-down JMJD6 expression were tested for STAT1 methylation, ISG activation, and HCV RNA. In vivo studies have been performed on C57Bl/6 mice (expressing HCV structural proteins or not) or chimeric mice with humanized livers fed control or ethanol diets. Results: AGS exposure to cells up-regulated JMJD6 expression in RLW cells. These results were corroborated by ethanol treatment of primary hepatocytes. The promethylating agent betaine reversed the effects of AGS/ethanol. Similar results were obtained in vivo, when mice were fed control/ethanol with and without betaine supplementation. Overexpression of JMJD6 suppressed STAT1 methylation, IFNα-induced ISG activation, and increased HCV-RNA levels. In contrast, JMJD6 silencing enhanced STAT1 methylation, ISG stimulation by IFNα and attenuated HCV-RNA expression in Huh7.5 cells. Conclusions: We conclude that arginine methylation of STAT1 is suppressed by JMJD6. Both HCV and acetaldehyde increase JMJD6 levels, thereby impairing STAT1 methylation and innate immunity protection in hepatocytes exposed to the virus and/or alcohol.

AB - Background & Aims: Alcohol-induced progression of hepatitis C virus (HCV) infection is related to dysfunction of innate immunity in hepatocytes. Endogenously produced interferon (IFN)α induces activation of interferon-stimulated genes (ISGs) via triggering of the Janus kinase–signal transducer and activator of transcription 1 (STAT1) pathway. This activation requires protein methyltransferase 1–regulated arginine methylation of STAT1. Here, we aimed to study whether STAT1 methylation also depended on the levels of demethylase jumonji domain-containing 6 protein (JMJD6) and whether ethanol and HCV affect JMJD6 expression in hepatocytes. Methods: Huh7.5-CYP (RLW) cells and hepatocytes were exposed to acetaldehyde-generating system (AGS) and 50 mmol/L ethanol, respectively. JMJD6 messenger RNA and protein expression were measured by real-time polymerase chain reaction and Western blot. IFNα-activated cells either overexpressing JMJD6 or with knocked-down JMJD6 expression were tested for STAT1 methylation, ISG activation, and HCV RNA. In vivo studies have been performed on C57Bl/6 mice (expressing HCV structural proteins or not) or chimeric mice with humanized livers fed control or ethanol diets. Results: AGS exposure to cells up-regulated JMJD6 expression in RLW cells. These results were corroborated by ethanol treatment of primary hepatocytes. The promethylating agent betaine reversed the effects of AGS/ethanol. Similar results were obtained in vivo, when mice were fed control/ethanol with and without betaine supplementation. Overexpression of JMJD6 suppressed STAT1 methylation, IFNα-induced ISG activation, and increased HCV-RNA levels. In contrast, JMJD6 silencing enhanced STAT1 methylation, ISG stimulation by IFNα and attenuated HCV-RNA expression in Huh7.5 cells. Conclusions: We conclude that arginine methylation of STAT1 is suppressed by JMJD6. Both HCV and acetaldehyde increase JMJD6 levels, thereby impairing STAT1 methylation and innate immunity protection in hepatocytes exposed to the virus and/or alcohol.

KW - Alcohol

KW - HCV

KW - JMJD6

KW - STAT1

UR - http://www.scopus.com/inward/record.url?scp=85042478712&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85042478712&partnerID=8YFLogxK

U2 - 10.1016/j.jcmgh.2017.10.004

DO - 10.1016/j.jcmgh.2017.10.004

M3 - Article

C2 - 29693039

AN - SCOPUS:85042478712

VL - 5

SP - 101

EP - 112

JO - Cellular and Molecular Gastroenterology and Hepatology

JF - Cellular and Molecular Gastroenterology and Hepatology

SN - 2352-345X

IS - 2

ER -